anti tyrp1 apc  (Novus Biologicals)

Journal: Communications Biology

Article Title: Pyroglutamation of cell surface proteins CD47 and TRP1 by glutaminyl cyclase modulates therapeutic antibody binding

doi: 10.1038/s42003-025-08938-4

Figure Lengend Snippet: a Crystal structure of human TRP1 (PDB: 5M8L), with the red motif indicating the N-terminal pGlu. b Titration of αTRP1 clone TA99 (1:1 dilution, commencing at 40 μg/mL) followed by αmouse IgG APC (1:800) on murine B16F10 wildtype (blue) and QPCTL KO (red) cells. Data represents individual data point and the line is the mean percentage of MFI of αTRP1 clone TA99 staining relative to the MFI of 40 μg/mL αTRP1 clone TA99 on wildtype cells (n = 3). c Flanvotumab (1:200) followed by αhuman clone QA19 (1:100) on B16F10 wildtype (blue) and QPCTL KO (red) cells. Data represents the percentage of MFI, + standard deviation error, bar of flanvotumab staining relative to the MFI flanvotumab on wildtype cells. One-way Anova was performed followed by the Dunnett’s multiple comparisons test (n = 3). d Representative histograms of αTRP1 clone TA99 (1:100) followed by αmouse IgG APC (1:800) or flanvotumab (1:200) followed by αhuman clone QA19 PE (1:100), unstained (gray) or secondary controls (dark gray) staining on B16F10 treated with 10 μM glutaminyl cyclase inhibitors PQ912 (red) or SEN177 (orange) or equal amounts of DMSO (blue) for three days. Bar graphs show the percentage of the MFI of αTRP1 clone TA99 or flanvotumab staining, + standard deviation error bar, relative to the MFI of DMSO control cells. One-way Anova was performed followed by the Dunnett’s multiple comparisons test (n = 3). e Representative histograms of αTRP1 clone TA99 (1:100) followed by αmouse IgG APC (1:800), or flanvotumab (1:50) followed by αhuman clone QA19 PE (1:100), unstained (gray) or secondary controls (dark gray) staining on primary immortalized human melanocytes treated with 10 μM glutaminyl cyclase inhibitors PQ912 (red) or SEN177 (orange) or equal amounts of DMSO (blue) for three days. Bar graphs show the percentage of the MFI of αTRP1 clone TA99 or flanvotumab, + standard deviation error bar, relative to the MFI of DMSO control cells. One-way Anova was performed followed by the Dunnett’s multiple comparisons test (n ≥ 3). Abbreviations: MFI, median fluorescent intensity; KO, knockout; US, unstained; Sec, secondary antibody only; PQ, PQ912; SEN, SEN177.

Article Snippet: Cells were incubated with polyclonal αTRP1 (1:100, # EPR13063 , Abcam) or αTRP1 clone TA99 (1:100, #SC-58438, Santa Cruz), diluted in 50 μL facs buffer supplemented with saponin for 30 minutes on ice shielded from light.

Techniques: Titration, Staining, Standard Deviation, Control, Knock-Out

Journal: Communications Biology

Article Title: Pyroglutamation of cell surface proteins CD47 and TRP1 by glutaminyl cyclase modulates therapeutic antibody binding

doi: 10.1038/s42003-025-08938-4

Figure Lengend Snippet: a K D calculation of TRP1 TA99 on B16F10 wildtype and QPCTL KO cells. ODE model fit for titration binding curves of WT and QPCTL KO conditions for the B16F10 cell line utilizing the full model in which K D values are estimated separately. b MFI of αTRP1 clone TA99 (1:50) followed by αmouse IgG APC (1:100) or αHIS PE (1:50) on grazoprevir treated HeLa cells expressing CAR-TRP1 in the presence or absence of 4 day treatment of 30 µM SEN177. Data represents the MFI, + standard deviation error bar. Unpaired T test was performed to assess the two groups (n = 3). c Percentage of CD8 + T-cells positive for the indicated markers that have been cocultured with B16F10 wildtype or QPCTL KO cells in the presence of 1, 0.1 or 0.01 ug/mL TRP1 TA99-CD3-bispecific antibody for 48 hours. Data represents the percentage, + standard deviation error bar. One-way Anova was performed to assess the groups (n = 3). Abbreviations: MFI, median fluorescent intensity; WT, wildtype; KO, knockout; PQ, PQ912; SEN, SEN177.

Article Snippet: Cells were incubated with polyclonal αTRP1 (1:100, # EPR13063 , Abcam) or αTRP1 clone TA99 (1:100, #SC-58438, Santa Cruz), diluted in 50 μL facs buffer supplemented with saponin for 30 minutes on ice shielded from light.

Techniques: Titration, Binding Assay, Expressing, Standard Deviation, Knock-Out

Journal: Journal for Immunotherapy of Cancer

Article Title: Cancer vaccines compensate for the insufficient induction of protective tumor-specific immunity of CD3 bispecific antibody therapy

doi: 10.1136/jitc-2024-010331

Figure Lengend Snippet: Combination of CD3xTRP1 with Fc-active TA99 augments primary survival but does not improve tumor-specific T-cell responses. ( A ) Treatment schedule for mice inoculated with B16F10 tumor cells and treated with a combination of CD3xTRP1 with TA99. ( B ) Kaplan-Meier survival graphs for indicated groups, numbers indicate surviving mice. ( C–D ) Expression of phenotypical markers on intratumoral macrophages ( C ), or cDC1s and cDC2s ( D ) treated according to the treatment scheme in . Numbers indicate average expression. Statistical significance is only shown for comparisons against the bsAb treatment group. ( E ) Tumor-specific responses on days 25 and 39 were measured by determining cytokine production in CD8 T cells from blood co-cultured with medium or IFNγ prestimulated B16F10 tumor cells. ( F ) Kaplan-Meier survival graphs for indicated groups, numbers indicate surviving mice. Data represented as mean on a heatmap for n=4–6 ( C–D , two untreated mice were excluded from the analysis based on tumor size<27 mm 3 ), or mean±SEM for n=3–14 ( E ). Statistics were calculated using Mantel-Cox log-rank tests ( B and F ), one-way ANOVA followed by Tukey’s post hoc tests to compare all treatments ( C–E ), or paired two-sided t-tests to compare different stimulations within the same treatment ( E ). ANOVA, analysis of variance; bsAb, bispecific antibody; cDC1s, conventional dendritic cells 1; cDC2s, conventional dendritic cells 2; IFNγ, interferon-gamma; iNOS, inducible nitric oxide synthase; TNFα, tumor necrosis factor alpha; TRP1, tyrosinase-related protein 1.

Article Snippet: Monospecific TA99 and Fc-silenced TA99 were generated in-house by Genmab by transfection of 293 F cells with expression vectors containing the relevant heavy and light chains.

Techniques: Expressing, Cell Culture